Critical Review of Formation and Growth of Anionic Vesicles Followed by Small-Angle Neutron Scattering, Grillo et

al., Langmuir 2003, 19, 4573-4581

Formation and Growth of Anionic Vesicles Followed by Small-Angle Neutron Scattering

(Langmuir 2003, 19, 4573-4581) by Grillo, Kats and Muratov attempts to study the kinetics behind
the formation of vesicles from micelles of surfactant AOT in aqueous salt solutions of varying
nature of ions (Na+, K+, Cl-, Br-) and concentrations using small angle neutron scattering (SANS).
They monitor the radius and the size distribution of vesicles using SANS. They then provide a
simple thermodynamic model to describe the mechanism of vesicle formation elucidating the fact
that vesicles are intermediates between micelles and bilayers. They follow up the thermodynamic
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argument with scaling arguments for growth of vesicles with time ( 6 ) and show that it
matches with their experimental observations. Understanding the stability and polydispersity of
the vesicles formed in such systems can provide guiding principles to model biological
membranes.
The system studied in the paper is the surfactant sodium bis(2-ethyl hexyl)sulfo-succinate
(AOT) in aqueous solutions of NaCl, NaBr, KCl, KBr in varying concentrations of surfactant and
electrolyte. The structure of micelles and vesicles formed have been observed using SANS on
the D22 instrument (Institut Laue Langevin, Grenoble). SANS provides information about the form
factor and the structure factor (if any) of a system. The contrast in SANS is obtained because of
difference in scattering length densities between various domains. In this paper they have used
data from elastic scattering from micellar solutions to determine the size of the vesicles formed.
A stopped flow apparatus was used to perform the experiment. Various volumes of the salt
solution were mixed with AOT solutions in H2O and D2O and passed through a stopped cell
where measurements were made with time. The data from SANS is in the form of I(q) v/s q, where
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I(q) is the intensity of scattering and q is the scattering wave vector given by, = sin ( ) where

2
is the wavelength of neutrons used, is the angle of scattered beam w.r.t. incident beam. Their
plots of I(q) v/s q showed an underlying spherical form factor for both micelles and vesicles but
no strong peaks implying the absence of an ordered structure. As one would expect vesicles have
peaks in the low q region whereas micelles are limited only to the high q because of smaller size.
The authors have provided a rigorous protocol for kinetic measurements of vesicles so to
obtain statistically relevant data. Two terms have been defined in the paper namely, tET or evolving
time which is time between diluting the surfactant solution and mixing it with salt solution and t DT
or dead time which is the minimum time after which a measurement is made after t ET. They
increased the acquisition time as a geometric progression and moved to lower for each sample
with passing time to account for the larger vesicles. These two measures insure confidence in the
measurement and the further interpretation.
Analysis of the I(q) v/s q plots obtained involves fitting the plot with a convolution of the
following functions: 1. A form factor(F(q,rext)) (spherical in our case); 2. Instrument resolution
function (R(q)); 3. Log normal distribution accounting for polydispersity in vesicles (G(r ext,)),
where is the standard deviation in sizes and rext the mean external radius of vesicles; 4. Linear
Background (IB). The model intensity thus becomes:

() = (, , ) ( , , ) 2 ( , ) +
0

Where Kc is the contrast for neutron scattering which is induced by them by using a mixture of
D2O and H2O to dissolve the AOT. The least squares error( 2 ) for the model is calculated to be
1.2 which seems to be an acceptable fit.
The authors argue that they can neglect scattering because of micelles in comparison of
vesicles for the following two reasons; first, the q range of interest for both of them is different
enough because of their large difference in sizes and second, the contrast provided by the heavy
water within vesicles is high enough that the scattering intensity from micelles is 10-100 times
lower than that of the vesicles. The arguments are acceptable and one can be confident of no
background micelle scattering in the observed form factor.
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Critical Review of Formation and Growth of Anionic Vesicles Followed by Small-Angle Neutron Scattering, Grillo et
al., Langmuir 2003, 19, 4573-4581

The authors proceed to prove the metastability of the vesicles with respect to micelles and
bilayers using real time measurements. Periodic scans were taken and two main features of the
scattering pattern were monitored and interpreted by the authors. First, the development and
growth of a plateau within a few milliseconds in the plot which implies increase in the concentration
and polydispersity of the micelles; second, a definite region of () 2 is seen over the course
of a few hours across decades of magnitude and is characteristic of bilayer form factor (lamellar).
This proves that vesicles are intermediates between micelles and bilayer which is insightful and
has been well reasoned out in the thermodynamic model presented in the paper. The authors
observe a decrease in speed of growth with increase in salt concentration and the extent of
deceleration followed the order KCl>KBr>NaCl>NaBr. The exact reason for this order has not
been made very clear in the paper which is one of its few negative points.
The theoretical treatment of the authors in the paper is simple and qualitatively accounts
for all experimental observations from SANS. They have been able to provide physical reasoning
for the metastability of the vesicles. In the literature the formation of initial smaller vesicles has
been argued on the increased entropy balancing the bend energy (which comes about because
of the different curvatures of the two concentric surfactant layers). The drive to form bilayers can
hence be easily understood so as to decrease this bending frustration. The authors argue that in
their system the ions from the salt shield the headgroups reducing the electrostatic repulsion
leading to the almost instantaneous vesicle formation.
They propose a multi-step process for the formation of mono-disperse vesicles (an
assumption based on the low value of obtained from SANS). The first fast step being the
screening of charge on the surfactants as discussed before. The second step involves the
insertion of a micelle into the vesicle making it grow. This step is repeated consecutively until the
free energy change of micelle insertion is equal to the chemical potential of a free micelle. This
picture of micelle insertion does make intuitive sense but requires actual visualization to prove it.
The authors propose a working model based on the above picture which does add credibility to
the picture, but does not completely prove it. The second assumption to make the model useful
is that the aggregation is driven by anharmonic contributions to bending elasticity (which appears
in the enthalpic term for free energy) as the harmonic contributions have no dependence on
aggregation number of the vesicles.
4 32 2
=
=
2

Furthermore, vesicle-vesicle interactions are assumed negligible and the rate of vesicle
growth is assumed to be controlled by micelle diffusion. These former assumption should hold for
the dilute solutions of AOT discussed in the paper. The later assumption subtly assumes that
micelle formation is instantaneous, which is true for these purposes as micelle formation happens
in the order of nanoseconds while the authors are interested in times upward of milliseconds. The
authors have overlooked clarifying these assumptions.
The authors then invoke the Landau kinetic theory setting up a direct propotionality of the
rate of growth of a vesicle to the total decrease in bending energy of the surface. The choice of
Landau theory was smart and in line with the assumption of monodisperse micelles and uniform
concentration of micelles and vesicles in the solution. A scaling analysis of the size of the vesicle
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yields = 6 where A is a constant. The change in R with t does not provide any insight to the
actual physical phenomenon involved in the formation of the vesicles, as to which might be the
rate limiting step; the diffusion of micelles or the insertion of micelles into a vesicle (the reaction).
To clarify this, the authors assume a Arrhenius type dependence of rate on T and a direct
correlation between the activation energy for the reaction and the square inverse of Debye length
( ), an assumption which is well understood by simple electrostatics; a smaller Debye length
implies a higher surface charge which would repel the insertion of a micelle into the vesicle thus
increasing the activation energy. Such an Arrhenius model should then lead to a exponential
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Critical Review of Formation and Growth of Anionic Vesicles Followed by Small-Angle Neutron Scattering, Grillo et
al., Langmuir 2003, 19, 4573-4581

dependence of R on Cs, which is not the case and hence the authors conclude that micelle
diffusion is the rate limiting step. This assumption seems incomplete in the sense that it does not
give a sense of how the micelle inserts, if it is chain by chain transfer or a distortion of the micelle
shape leading to a key in the lock kind of insertion or some other sort. The authors fail to address
this issue in the paper notwithstanding the difficulty in actually determining the mechanism. On
the other hand the authors do mention other vesicles growth processes from the literature leading
to other scaling laws. Each of the other methods assume a different model of growth leading to a
different analysis.
The major drawback of the paper is in their assumption of mono disperse micelles and the
development of the model based on that. The finite , as they find experimentally, leads to a
variation in the exponent in the scaling obtained by fitting especially at longer times where the
polydispersity of the vesicles is more predominant. The authors try to add physical significance to
the pre-factor A but end up giving mathematical arguments for its concentration dependence.
Using the Debye approximation for surface potential, the authors determine that A has an inverse
(1/6)th power concentration dependence, which is coincidentally equal R-t scaling.
In summary, the paper has done a very good job of proposing a thermodynamic and kinetic
hypothesis and validating it experimentally with SANS experiments. The simplicity of the model
is singularly amazing. The authors made clear the different variables in the AOT-water-salt system
and varied each variable separately to yield a clear picture. The SANS experiments were also
well engineered in the stop-flow cell and especially with acquisition times so as to obtain
statistically relevant data. Having said that, the fitting of the plots was done on fewer points than
required to make one feel comfortable. In addition to that the authors do not validate the
applicability of the Landau theory for describing the kinetics of vesicle growth. Furthermore some
the assumptions made in the paper require a few lines of reasoning so as to make the paper a
smooth read. Overall the paper was able to propose a mechanism for vesicle growth, validate it
with experiments and also observe its variation with added salt and changing surfactant
concentration. The authors provide scope of further research in this area by changing surfactants,
solvents and the nature of salts to strengthen their hypothesis. The paper also does a good job
of opening up a number of avenues related to studying kinetics of self-assembly of surfactants
into liquid crystalline phases using SANS.

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